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Liquid chromatography reagent for determining glycosylated hemoglobin and preparation method of liquid chromatography reagent

A glycated hemoglobin and liquid chromatography technology, applied in the field of medical testing, can solve the problems of unguaranteed accuracy and precision, expensive equipment, long testing time, etc. Stability and precision, effects of improving specificity and stability

Inactive Publication Date: 2021-03-12
ZYBIO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the ion-exchange HPLC method is a standard reference method guided by IFCC, which has the advantages of good precision and accurate detection results. Usually, the precision CV value detected by the ion-exchange HPLC method is less than 3%, but the required equipment is expensive and the detection cost is high. , and its detection takes a long time
The opposite is immunochromatography, which is easy to carry out in primary hospitals because of its simple operation, but due to its limitations and inhomogeneity of chromatographic materials, the precision and accuracy of the test results are not good. Immunochromatography is usually used The precision CV value of chromatography detection is less than 15%, and the accuracy and precision of detection cannot be guaranteed

Method used

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  • Liquid chromatography reagent for determining glycosylated hemoglobin and preparation method of liquid chromatography reagent
  • Liquid chromatography reagent for determining glycosylated hemoglobin and preparation method of liquid chromatography reagent
  • Liquid chromatography reagent for determining glycosylated hemoglobin and preparation method of liquid chromatography reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 Preparation of glycosylated hemoglobin detection test strip

[0036] 1. Preparation of Reagent 1

[0037] Configure according to the following formula, stir and mix thoroughly, and store at 2-8°C.

[0038]

[0039] The remaining solvent is purified water

[0040] 2. Preparation of Reagent 2

[0041] Configure according to the following formula, stir and mix thoroughly, and store at 2-8°C.

[0042]

[0043] The remaining solvent is purified water

[0044] 3. Assembly of test strips

[0045] (1) Treatment of nitrocellulose membrane

[0046] a. Preparation of test line

[0047] Dilute the anti-human glycosylated hemoglobin HbA1c monoclonal antibody to 2.0mg / ml with 20mmol / L PH7.2 PBS buffer, and draw a detection line at the left end of the nitrocellulose membrane with an amount of 0.6ul / cm.

[0048] b. Preparation of quality control line

[0049] Draw a quality control line on the right end of the nitrocellulose membrane with the goat anti-genus ...

Embodiment 2

[0056] The using method of embodiment 2 test strips

[0057] (1) Add 0.05ml of the sample to be tested into 2.45ml of reagent R1, stir and mix to complete rapid hemolysis;

[0058] (2) Mix the mixed solution in step (1) with 0.1ml of reagent R2, stir and mix, so that the hemoglobin antibody in the sample and the hemoglobin and anti-human hemoglobin antibody fully react;

[0059] (3) Add dropwise 60ul of the mixed solution to the sample hole of the test strip prepared in Example 1;

[0060] (4) Chromatography of the sample mixture until the detection line of the test strip binds to the anti-human glycosylated hemoglobin antibody, and then chromatographs until the quality control line binds to the goat anti-mouse antibody;

[0061] (5) Measure the gray value after the reaction with an immunoquantitative analyzer (Chongqing Zhongyuan Huiji Biotechnology Co., Ltd., Q8pro), and calculate the proportion of glycosylated hemoglobin in the sample according to the gray value.

Embodiment 3

[0062] Embodiment 3 performance test

[0063] (1) Test results of HbA1c standard products

[0064] Adopt the method for using the test strip in Example 2, select the HbA1c standard substance of different concentrations (get seven different concentrations, be respectively 2%, 4%, 6%, 8%, 12%, 14%, 16% for use) %, each concentration was set to 3 repetitions), the samples to be tested after the pretreatment of reagents R1 and R2 were respectively dropped onto the test strip sample pad prepared in Example 1, and the immunoquantitative analyzer (Chongqing Zhongyuan Huiji Biotechnology Co., Ltd., Q8pro) to read the signal. The test results are shown in the table below:

[0065] Table 1 HbA1c standard test results

[0066]

[0067] Take the HbA1c antigen standard substance concentration as the abscissa, and the measured average concentration as the ordinate, establish a logarithmic equation and fit the standard curve, and the results are shown in the attached figure 2 As show...

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Abstract

The invention discloses a liquid chromatography reagent for determining glycosylated hemoglobin. The liquid chromatography reagent comprises an immunochromatography test strip, a reagent R1 and a reagent R2; the immunochromatography test strip comprises a bottom lining, a sample pad, a nitrocellulose membrane and a water absorption pad; one or more detection lines are fixed on the nitrocellulose membrane; the detection line is an anti-human glycosylated hemoglobin antibody; the reagent R1 contains 110g / L of SDS; and the reagent R2 contains a tracer particle labeled anti-human hemoglobin antibody. According to the liquid chromatography reagent for determining the glycosylated hemoglobin and the preparation method of the liquid chromatography reagent of the present invention, the special reagent R2 is prepared, the tracer particle labeled antibody is dissolved in the reagent R2; and the reagent R2 is combined with the tracer particle labeled antibody in the pre-treatment stage of a sample to be detected, and the reagent R1 is prepared so as to carry out hemolysis treatment on the sample in the pre-treatment stage; on one hand, the preparation process of the test strip is simplified,and on the other hand, the specificity and stability of the test strip are improved, so that the detection precision CV value is lower than 3%.

Description

technical field [0001] The invention relates to the field of medical testing, in particular to a liquid chromatography reagent for measuring glycosylated hemoglobin and a preparation method thereof Background technique [0002] Glycosylated hemoglobin HbA1c is the product of the combination of hemoglobin in red blood cells and sugar in serum. It is formed through a slow, continuous and irreversible glycation reaction, and its content depends on the blood sugar concentration and the contact time between blood sugar and hemoglobin. It is related to the survival of red blood cells, and its synthesis rate is proportional to the amount of red blood cell glycation in the average 120d life cycle. , clinically, HbA1c represents the control of the average blood sugar level of the human body in the last 2-3 months, and has nothing to do with factors such as blood draw time, whether the patient is fasting, and whether insulin is used. Therefore, HbA1c is the gold standard for measuring...

Claims

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Application Information

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IPC IPC(8): G01N33/72G01N33/577G01N33/558G01N33/543G01N33/532
CPCG01N33/723G01N33/577G01N33/558G01N33/54313G01N33/532
Inventor 肖江涛秦川
Owner ZYBIO INC